Mechanisms of Human Erythrocytic Bioactivation of Nitrite

Liu, Chen; Wajih, Nadeem; Liu, Xiaohua; Basu, Swati; Janes, John; Marvel, Madison; Keggi, Christian; Helms, Christine; Lee, Amber N.; Belanger, Andrea; Diz, Debra I.; Laurienti, Paul J.; Caudell, David L.; Wang, Jun; Gladwin, Mark T.; Kim‐Shapiro, Daniel B.

doi:10.1074/jbc.m114.609222

Cited by 70 publications

(86 citation statements)

References 47 publications

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“…5B). The observed changes in platelet activation by human RBCs and nitrite are significant (P = 0.038) and consistent with previous results (11) (Fig. S4).…”

Section: Resultssupporting

confidence: 92%

“…5). We observed that nitrite inhibits platelet activation in the presence of human deoxyHb as previously reported (11,13) (Fig. 5A).…”

Section: Resultssupporting

confidence: 89%

“…3D shows representative trace signals of NO production. The injection of Hb caused NO release as described in previous studies (11,23). However, the injection of deoxyGbX resulted in a greater release of NO into the gas phase compared with human deoxyHb.…”

Section: Resultsmentioning

confidence: 52%

“…In particular, Mb is the main regulator of the vasodilatory response in muscles (27) and Hb appears to be the main protein regulating NO production from nitrite in the human RBCs (11). As platelet activation is inhibited by NO, platelet activation assays are useful and biologically relevant models to assess the release of NO from nitrite reactions with globins and intact RBCs.…”

Section: Resultsmentioning

confidence: 99%

“…sGC activation catalyzes the formation of the second messenger cGMP and subsequent inhibition of intracellular calcium release, resulting in decreased platelet aggregation. Recent studies from multiple laboratories show that low concentrations of nitrite in the presence of RBCs inhibit human platelets activation in vitro and in vivo in mice and humans (10)(11)(12). It has been demonstrated that in low oxygen conditions, Hb functions as a nitrite reductase, generating NO from nitrite and thus producing an antiaggregation effect on platelets (13,14).…”

mentioning

confidence: 99%

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Globin X is a six-coordinate globin that reduces nitrite to nitric oxide in fish red blood cells

Cortí

Xue

Tejero

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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The discovery of novel globins in diverse organisms has stimulated intense interest in their evolved function, beyond oxygen binding. Globin X (GbX) is a protein found in fish, amphibians, and reptiles that diverged from a common ancestor of mammalian hemoglobins and myoglobins. Like mammalian neuroglobin, GbX was first designated as a neuronal globin in fish and exhibits six-coordinate heme geometry, suggesting a role in intracellular electron transfer reactions rather than oxygen binding. Here, we report that GbX to our knowledge is the first six-coordinate globin and the first globin protein apart from hemoglobin, found in vertebrate RBCs. GbX is present in fish erythrocytes and exhibits a nitrite reduction rate up to 200-fold faster than human hemoglobin and up to 50-fold higher than neuroglobin or cytoglobin. Deoxygenated GbX reduces nitrite to form nitric oxide (NO) and potently inhibits platelet activation in vitro, to a greater extent than hemoglobin. Fish RBCs also reduce nitrite to NO and inhibit platelet activation to a greater extent than human RBCs, whereas GbX knockdown inhibits this nitrite-dependent NO signaling. The description of a novel, sixcoordinate globin in RBCs with dominant electron transfer and nitrite reduction functionality provides new insights into the evolved signaling properties of ancestral heme-globins.nitrite | nitric oxide | blood | RBC | platelet

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“…5B). The observed changes in platelet activation by human RBCs and nitrite are significant (P = 0.038) and consistent with previous results (11) (Fig. S4).…”

Section: Resultssupporting

confidence: 92%

“…5). We observed that nitrite inhibits platelet activation in the presence of human deoxyHb as previously reported (11,13) (Fig. 5A).…”

Section: Resultssupporting

confidence: 89%

Section: Resultsmentioning

confidence: 52%

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Globin X is a six-coordinate globin that reduces nitrite to nitric oxide in fish red blood cells

Cortí

Xue

Tejero

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Humoral Mediators of Remote Ischemic Conditioning: Important Role of eNOS/NO/Nitrite

Hess

Hoda

Khan

2016

Acta Neurochirurgica Supplement

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Remote ischemic conditioning (RIC) is a powerful cardioprotectant and neuroprotectant. The mechanism of protection likely involves circulating, blood-borne mediators that transmit the signal from the periphery to the brain. The neuroprotective effect of RIC may be partially related to improvements in cerebral blood flow (CBF). Nitrite is a key circulating mediator of RIC and may be a mediator of increased CBF and also mediate cytoprotection through its effects on nitrosylation of mitochondrial proteins such as complex I. Measuring plasma nitrite may serve as an important blood biomarker, and measuring CBF by techniques such as MRI arterial spin labeling (ASL) may be an ideal surrogate imaging biomarker in clinical trials of RIC.

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Discovery and microassay of a nitrite-dependent carbonic anhydrase activity by stable-isotope dilution gas chromatography–mass spectrometry

et al. 2015

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The intrinsic activity of carbonic anhydrase (CA) is the hydration of CO2 to carbonic acid and its dehydration to CO2. CA may also function as esterase and phosphatase. Recently, we demonstrated that renal CA is mainly responsible for the reabsorption of nitrite (NO2(-)) which is the most abundant reservoir of the biologically highly potent nitric oxide (NO). By means of a stable-isotope dilution GC-MS method, we discovered a novel CA activity which strictly depends upon nitrite. We found that bovine erythrocytic CAII (beCAII) catalyses the incorporation of (18)O from H2 (18)O into nitrite at pH 7.4. After derivatization with pentafluorobenzyl bromide, gas chromatographic separation and mass spectrometric analysis, we detected ions at m/z 48 for singly (18)O-labelled nitrite ((16)O=N-(18)O(-)/(18)O=N-(16)O(-)) and at m/z 50 for doubly (18)O-labelled nitrite ((18)O=N-(18)O(-)) in addition to m/z 46 for unlabelled nitrite. Using (15)N-labelled nitrite ((15)NO2 (-), m/z 47) as an internal standard and selected-ion monitoring of m/z 46, m/z 48, m/z 50 and m/z 47, we developed a GC-MS microassay for the quantitative determination of the nitrite-dependent beCAII activity. The CA inhibitors acetazolamide and FC5 207A did not alter beCAII-catalysed formation of singly and doubly (18)O-labelled nitrite. Cysteine and the experimental CA inhibitor DIDS (a diisothiocyanate) increased several fold the beCAII-catalysed formation of the (18)O-labelled nitrite species. Cysteine, acetazolamide, FC5 207A, and DIDS by themselves had no effect on the incorporation of (18)O from H2 (18)O into nitrite. We conclude that erythrocytic CA possesses a nitrite-dependent activity which can only be detected when nitrite is used as the substrate and the reaction is performed in buffers of neutral pH values prepared in H2 (18)O. This novel CA activity, i.e., the nitrous acid anhydrase activity, represents a bioactivation of nitrite and may have both beneficial (via S-nitrosylation and subsequent NO release) and possibly adverse (via C- and N-nitrosylation) effects in living organisms.

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Mechanisms of Human Erythrocytic Bioactivation of Nitrite

Cited by 70 publications

References 47 publications

Globin X is a six-coordinate globin that reduces nitrite to nitric oxide in fish red blood cells

Globin X is a six-coordinate globin that reduces nitrite to nitric oxide in fish red blood cells

Humoral Mediators of Remote Ischemic Conditioning: Important Role of eNOS/NO/Nitrite

Discovery and microassay of a nitrite-dependent carbonic anhydrase activity by stable-isotope dilution gas chromatography–mass spectrometry

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